Project Summary The laboratories of Drs. Alland, Perlin, Dick, Dartois, Freundlich, Kreiswirth and Neiditch have been working collaboratively for many years, conducting cutting edge research in the infectious disease field. One of the major focuses of these laboratories is to discover and develop new antimicrobial drugs against high-threat pathogens such as drug-resistant Mycobacterium tuberculosis, nontuberculous mycobacterial (NTM) disease, select agents and drug resistant bacterial and fungal pathogens. These laboratories are also global leaders in developing advanced PCR based molecular diagnostic assays. Additional NIH funded projects include studies of M. tuberculosis biomarkers that predict poor treatment outcomes. All of these projects rely on extensive characterization of interactions between bacteria and antibiotics, or complex studies of drug properties. The purpose of this grant is to acquire an Echo 650 Liquid Handler (Labcyte Inc.) to support a massive scale-up and an increase in the complexity of bacteria-drug and drug-ADME studies by these investigators and other researchers at Rutgers University and at the Hackensack Meridian Health Center for Discovery & Innovation. This liquid handler rapidly and accurately dispenses compounds, chemicals and reagents in 2.5 nL packets using sonic waves without using any type of tips or cartridges. This process permits rapid construction of complex multi-reagent dilution series in plates at densities as high 3,456 wells, and eliminates inaccuracies caused by liquid adherence to pipette tips and the resulting carryover common to all classical serial dilution approaches. We have also developed an application that enables bacterial dilution series that is as accurate as reagent dilutions. This instrument permits experiments that would otherwise be completely impractical or not worth pursuing due to pipetting inaccuracies. It reduces preparation times from hours to minutes, dramatically increases reproducibility, and produces substantial savings in plastic consumables. Based on acquisition of this equipment we aim to perform 1) Fast high-throughput minimal inhibitory concentration (MIC) drug screening of a wide-range of anti-microbial compounds against M. tuberculosis, NTMs, fungi and other high priority bacteria. 2) Multi-drug combination studies to test for drug synergy to determine in vitro efficacy of multi-drug regimens. 3) Large scale susceptibility testing of M. tuberculosis clinical isolates to TB drugs using fractional dilutions. 4) Achieve a 100-fold reduction in the cost of preparing Illumina DNA Flex libraries for whole genome sequencing and RNAseq by performing nano-scale library generation. 5) Develop and perform large scale plate based enzymatic and cell based assays. 6) Develop new applications in the field of antimicrobial compound discovery and routinely run otherwise complex and time intensive multi-analyte analyses. We have evaluated other liquid handlers but none of them fully meet our application needs except Echo series. In conclusion, integration of the Echo 650 into our existing studies will revolutionize the quantity and quality of our data production, enabling us to dramatically expand the scope and depth of our NIH funded projects.